Maturation of porcine tracheal cholinergic and inhibitory non-adrenergic, non-cholinergic innervation.

Abstract

It is known that both excitatory cholinergic neural activity and nitric oxide (NO) release from inhibitory non-adrenergic, non-cholinergic (iNANC) nerves are important determinants of adult human airway smooth muscle tone. However, the level of cholinergic and iNANC activity in the newborn is unknown. Therefore, isolated trachealis muscle strips were obtained from neonatal, 10-day-old, and adult pigs. To quantify cholinergic nerve activity, isometric contractions were elicited by stimulating the nicotinic receptors of the cholinergic intramural ganglia with dimethyl-phenyl-piperazinium (DMPP). After ganglionic blockade with hexamethonium, the post-ganglionic cholinergic nerves were subjected to electrical field stimulation (EFS). To assess whether DMPP and EFS were also stimulating nitric oxide release from iNANC nerves, the stimulations were repeated after NO-synthase inhibition with 3 x 10(-5) M NG-monomethyl-L-arginine acetate (LNMMA). All responses were expressed as a percentage of maximal response to 10(-4) M acetylcholine. Although no significant age-related differences in the contractile responses to DMPP were demonstrated between the three age groups, the EFS responses were significantly different. At EFS frequencies of 1 and 2 Hz the responses of all three groups were different (P < 0.05, unpaired t-test). At frequencies of 0.125 and 0.5 Hz the contractile responses in the neonatal and 10-day-old groups were less than in the adult animal tissues (P < 0.05, unpaired t-test). At 4 Hz and above, the responses of the 10-day-old and adult groups became greater than the neonatal (P < 0.05, unpaired t-test). LNMMA did not significantly increase the contractile responses to DMPP at any age. Only in the neonatal rat and 10-day-old groups was a significant increase of the EFS frequency responses by LNMMA demonstrated (140.4 +/- 38.2% and 144.2 +/- 41.2%, respectively, at 0.5 Hz, ANOVA, P < 0.001). In conclusion, the DMPP responses suggest that the neonatal porcine cholinergic innervation is mature at birth. This contrasts with a frequency-dependent increase in EFS responses from birth. This may be due in part to a relative increase in iNANC modulation of post-ganglionic cholinergic activity at birth. This mechanism is not responsible for the increase in higher EFS frequency responses with post-natal maturation. Inhibitory iNANC activity may serve a protective role in the newborn, given the reported hyper-responsiveness of airway smooth muscle, small airway caliber, and high chest wall compliance at this age.